Mscarlet 1

Manufactured by Addgene

MScarlet-I is a fluorescent protein that emits light in the red region of the visible spectrum. It can be used as a reporter or marker in biological experiments.

Automatically generated - may contain errors

Lab products found in correlation

Opti mem, by Thermo Fisher Scientific (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Memerald lifeact, by Addgene (1 mentions) Memerald, by Addgene (1 mentions) Pcw cas9, by Addgene (1 mentions) Pmcb320, by Addgene (1 mentions) Pmj179, by Addgene (1 mentions) Mirfp670, by Addgene (1 mentions) Pgex 6p 1, by GE Healthcare (1 mentions) Pdonr221, by Thermo Fisher Scientific (1 mentions)

2 protocols using mscarlet 1

Fluorescent Protein Transfection Protocol

Cited 1 time

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FusionRed-F-tractin has been previously described [3 (link)]. EGFP-F-tractin (no. 58473), mEmerald (no. 53975), LifeAct-mEmerald (no. 54148), mScarlet-I (no. 85044), and LifeAct-mScarlet-I (no. 85054) were obtained from Addgene (Watertown, MA). 1 μg of plasmid was used to transfect 750,000 cells in each well of a 6-well plate using Lipofectamine 2000 (5 μL; Thermo Fisher Scientific) in OptiMEM (400 μL; Thermo Fisher Scientific). After 30 min at room temperature, plasmid in Lipofectamine 2000/OptiMEM was then incubated with cells in complete media (2 mL) overnight.

Vosatka K.W., Lavenus S.B, & Logue J.S. (2022). A novel Fiji/ImageJ plugin for the rapid analysis of blebbing cells. PLoS ONE, 17(4), e0267740.

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Cloning and Expression of RAB34 Constructs

Cited 2 times

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Gibson assembly, Gateway cloning, and standard molecular biology were used for DNA cloning. Human RAB34 cDNA was obtained from GE Healthcare (MHS6278-202807876) and cloned into Gateway pENTR plasmid pDONR221 (Invitrogen); RAB34 mutants were prepared by site-directed mutagenesis of pENTR-Rab34. Plasmids for tetracycline-inducible expression of LAP-Rab34 (LAP consists of GFP, TEV protease site, and S-tag) were constructed by modification of pCW-Cas9 (Addgene #50661, gift from Eric Lander and David Sabatini). Stable expression of fluorescently tagged ciliary and centriolar proteins was achieved by modification of pCW-Cas9 using cDNAs for the PGK promoter, ARL13B (Addgene #40879, gift from Tamara Caspary), EHD1 (gift from Chris Westlake), Rab8 (gift from Maxence Nachury), and Centrin2 (gift from Tim Stearns), miRFP-670 (Addgene #79987, gift from Vladislav Verkhusha), and mScarlet-I (Addgene #85044, gift from Dorus Gadella). Plasmids for CRISPR-based knockout were constructed using pMCB320 (Addgene #89359, gift from Michael Bassik) and pMJ179 (Addgene #89556, gift from Jonathan Weissman); see Key Resources Table for sgRNA sequences. Plasmids for transient transfection of RAB34 were made by Gateway cloning using pEF5-FRT-LAP-DEST^{55 (link)},^{56 (link)}; for bacterial protein expression, RAB34 variants were cloned into pGEX-6P1 (GE Healthcare).

Ganga A.K., Kennedy M.C., Oguchi M.E., Gray S., Oliver K.E., Knight T.A., De La Cruz E.M., Homma Y., Fukuda M, & Breslow D.K. (2021). Rab34 GTPase mediates ciliary membrane formation in the intracellular ciliogenesis pathway. Current biology : CB, 31(13), 2895-2905.e7.

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